Open AccessCloning and characterization of the Halobacillus trueperi betH gene, encoding the transport system for the compatible solute glycine betaine
Author(s) -
Lu Weidong,
Zhao Baisuo,
Feng Deqin,
Yang Susheng
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2004.tb09615.x
Subject(s) - betaine , microbiology and biotechnology , biology , bacillus subtilis , gene , southern blot , molecular cloning , biochemistry , genetics , peptide sequence , bacteria
Halobacillus trueperi accumulates glycine betaine under condition of high osmolarity. A fragment of the glycine betaine transporter betH gene was obtained from the genome of H. trueperi with degenerate primers. Through Southern blot hybridization and inverse PCR, a 5.1 kb Eco RI fragment containing the complete betH gene was identified and subsequently sequenced. The betH gene was predicted to encode a 55.2 kDa protein (504 amino acid residues) with 12 transmembrane regions. BetH showed 56% identity to the OpuD of Bacillus subtilis which belongs to the betaine/carnitine/choline transporter (BCCT) family. Its putative promoter region was highly homologous to σ B ‐dependent promoter of B. subtilis . A 2.6 kb fragment containing the betH gene was cloned into pUC18 and transformed into the Escherichia coli MKH13. The accumulation of glycine betaine in transformed E. coli MKH13 bacteria was confirmed using 13 C nuclear magnetic resonance spectroscopy.